Movement of soil-applied imidacloprid and thiamethoxam into nectar and pollen of squash (Cucurbita pepo)

There has been recent interest in the threat to bees posed by the use of systemic insecticides. One concern is that systemic insecticides may translocate from the soil into pollen and nectar of plants, where they would be ingested by pollinators. This paper reports on the movement of two such systemic neonicotinoid insecticides, imidacloprid and thiamethoxam, into the pollen and nectar of flowers of squash (Cucurbita pepo cultivars "Multipik," "Sunray" and "Bush Delicata") when applied to soil by two methods: (1) sprayed into soil before seeding, or (2) applied through drip irrigation in a single treatment after transplant. All insecticide treatments were within labeled rates for these compounds. Pollen and nectar samples were analyzed using a standard extraction method widely used for pesticides (QuEChERS) and liquid chromatography mass spectrometric analysis. The concentrations found in nectar, 10 ± 3 ppb (mean ± s.d) for imidacloprid and 11 ± 6 ppb for thiamethoxam, are higher than concentrations of neonicotinoid insecticides in nectar of canola and sunflower grown from treated seed, and similar to those found in a recent study of neonicotinoids applied to pumpkins at transplant and through drip irrigation. The concentrations in pollen, 14 ± 8 ppb for imidacloprid and 12 ± 9 ppb for thiamethoxam, are higher than those found for seed treatments in most studies, but at the low end of the range found in the pumpkin study. Our concentrations fall into the range being investigated for sublethal effects on honey bees and bumble bees.     

Potential risks of systemic imidacloprid to parasitoid natural enemies of a cerambycid attacking Eucalyptus

A community of insect herbivores has established on eucalyptus species in California following their introduction from Australia. A number of the species are under complete or partial biological control. A response to introduction of additional pest species into the complex has been the application of systemic insecticides to infested trees. Natural enemies that have been introduced to control the various pest species feed on the nectar of treated trees, and thus may be affected by these pesticides. In this study, Eucalyptus rudis trees were treated at label rates with the neonicotinoid systemic insecticide imidacloprid and nectar sampled at 5 months post-treatment during the spring bloom. The concentration of imidacloprid and its toxic metabolites in nectar was measured by ELISA at 660 ppb. Adults of the encyrtid egg parasitoid Avetianella longoi that were fed floral nectar collected from treated trees had significantly lower survival and reproductive fitness than adults fed nectar from untreated trees. In feeding bioassays, in which the adults were fed a range of concentrations of imidacloprid in sugar water, the LC₅₀ for A. longoi was 212 ppb imidacloprid. Bioassays were also conducted with the braconid larval parasitoid, Syngaster lepidus. The LC₅₀ for S. lepidus was 288 ppb imidacloprid. The insecticide appears to be concentrating in the nectar at higher levels than reported from other plant species and at concentrations exceeding the LC₅₀ for two important parasitoids. If tree treatments become widespread as a result of continual introductions of new eucalypt herbivores, established biological control programs could be at significant risk.     

Imidacloprid Alters Foraging and Decreases Bee Avoidance of Predators

Concern is growing over the effects of neonicotinoid pesticides, which can impair honey bee cognition. We provide the first demonstration that sublethal concentrations of imidacloprid can harm honey bee decision-making about danger by significantly increasing the probability of a bee visiting a dangerous food source. Apis cerana is a native bee that is an important pollinator of agricultural crops and native plants in Asia. When foraging on nectar containing 40 µg/L (34 ppb) imidacloprid, honey bees (Apis cerana) showed no aversion to a feeder with a hornet predator, and 1.8 fold more bees chose the dangerous feeder as compared to control bees. Control bees exhibited significant predator avoidance. We also give the first evidence that foraging by A. cerana workers can be inhibited by sublethal concentrations of the pesticide, imidacloprid, which is widely used in Asia. Compared to bees collecting uncontaminated nectar, 23% fewer foragers returned to collect the nectar with 40 µg/L imidacloprid. Bees that did return respectively collected 46% and 63% less nectar containing 20 µg/L and 40 µg/L imidacloprid. These results suggest that the effects of neonicotinoids on honey bee decision-making and other advanced cognitive functions should be explored. Moreover, research should extend beyond the classic model, the European honey bee (A. mellifera), to other important bee species.     

Soil-applied imidacloprid is translocated to nectar and kills nectar-feeding Anagyrus pseudococci (Girault) (Hymenoptera: Encyrtidae)

Behavior was altered and survivorship was reduced when parasitoids, Anagyrus pseudococci (Girault) (Hymenoptera: Encyrtidae), were fed flowers from buckwheat, Fagopyrum esculentum L. (Polygonaceae), treated with soil applications of imidacloprid (Marathon 1% G). Parasitoids at 1 d had significantly reduced survivorship of 38 +/- 6.7% on label rate and 17 +/- 4.2% on twice label rate compared with 98 +/- 1.2% on untreated flowers. Parasitoids trembled 88% on label rate and 94% on twice label rate compared with 0% on untreated flowers. Residue analysis on a composite sample of 425 flowers showed that imidacloprid concentration was 6.6 +/- 1.0 ppm (16 ppb/flower) in label rate, 12.3 +/- 2.7 ppm (29 ppb/flower) in twice label rate, and 0 ppb in untreated flowers. The hydroxy metabolite concentration was 1.1 ppm (2.4 ppb/flower) in label rate, 1.9 ppm (4.4 ppb/flower) in twice label rate, and 0 ppm in untreated flowers. The olefin metabolite concentration was 0.2 ppm (0.5 ppb/flower) in label rate, 0.5 ppm (1.1 ppb/flower) in twice label rate, and 0 ppm in untreated flowers. Soil-applied imidacloprid used at flowering may be translocated to nectar in higher concentration compared with the imidacloprid seed treatment Gaucho. Considerable research has studied effects of Gaucho-treated canola, sunflower, and maize on behavior and mortality of Apis mellifera L. In our laboratory, we showed that translocation of imidacloprid to flowers reduced survivorship and altered behavior of pink lady beetle, Coleomegilla maculata DeGeer (Smith and Krischik 1999) and green lacewing, Chrysoperla carnea Stephens (Rogers et al. 2007).     

Nontarget impacts of neonicotinoids on nectar-inhabiting microbes

Plant-systemic neonicotinoid (NN) insecticides can exert non-target impacts on organisms like beneficial insects and soil microbes. NNs can affect plant microbiomes, but we know little about their effects on microbial communities that mediate plant-insect interactions, including nectar-inhabiting microbes (NIMs). Here we employed two approaches to assess the impacts of NN exposure on several NIM taxa. First, we assayed the in vitro effects of six NN compounds on NIM growth using plate assays. Second, we inoculated a standardised NIM community into the nectar of NN-treated canola (Brassica napus) and assessed microbial survival and growth after 24 h. With few exceptions, in vitro NN exposure tended to decrease bacterial growth metrics. However, the magnitude of the decrease and the NN concentrations at which effects were observed varied substantially across bacteria. Yeasts showed no consistent in vitro response to NNs. In nectar, we saw no effects of NN treatment on NIM community metrics. Rather, NIM abundance and diversity responded to inherent plant qualities like nectar volume. In conclusion, we found no evidence that NIMs respond to field-relevant NN levels in nectar within 24 h, but our study suggests that context, specifically assay methods, time and plant traits, is important in assaying the effects of NNs on microbial communities.     

Repression and Recuperation of Brood Production in Bombus terrestris Bumble Bees Exposed to a Pulse of the Neonicotinoid Pesticide Imidacloprid

Currently, there is concern about declining bee populations and some blame the residues of neonicotinoid pesticides in the nectar and pollen of treated crops. Bumble bees are important wild pollinators that are widely exposed to dietary neonicotinoids by foraging in agricultural environments. In the laboratory, we tested the effect of a pulsed exposure (14 days ‘on dose’ followed by 14 days ‘off dose’) to a common neonicotinoid, imidacloprid, on the amount of brood (number of eggs and larvae) produced by Bombus terrestris L. bumble bees in small, standardised experimental colonies (a queen and four adult workers). During the initial ‘on dose’ period we observed a dose-dependent repression of brood production in colonies, with productivity decreasing as dosage increased up to 98 µg kg⁻¹ dietary imidacloprid. During the following ‘off dose’ period, colonies showed a dose-dependent recuperation such that total brood production during the 28-day pulsed exposure was not correlated with imidacloprid up to 98 µg kg⁻¹. Our findings raise further concern about the threat to wild bumble bees from neonicotinoids, but they also indicate some resilience to a pulsed exposure, such as that arising from the transient bloom of a treated mass-flowering crop.     

Impact of flower rewards on phytophagous insects: importance of pollen and nectar for the development of the pollen beetle (<Emphasis Type="Italic">Brassicogethes aeneus</Emphasis>)

Entomophilous plants reward pollinators with provision of nutrient-rich foods such as pollen and nectar. These rewards contain compounds that are essential to insect development and can be used by pollinators as well as herbivorous insects. The pollen beetle (Brassicogethes aeneus, syn. Meligethes aeneus) whose larvae develop in oilseed rape flowers (Brassica napus) is known to feed on pollen. Previous studies already showed the importance of pollen on the development of this insect but it seems that other resource, such as nectar, could also be used. The purpose of this study was to assess the respective roles of pollen and nectar on pollen beetle development. We tested their role with behavioural and developmental experiments using flowers where the presence and absence of nectar and pollen varied. Larvae, irrespective of their instar, fed both on anthers and nectar. Nectar did not influence larval development or adult survival while pollen influenced development by increasing both larval and adult weight. However, pollen did not affect larval or adult survival nor development time. These results indicate that pollen beetle larvae are adapted to deal with various diets and can complete their development without pollen or nectar.     

In search of the best pollinators for canola (<Emphasis Type="Italic">Brassica napus</Emphasis> L.) production in Pakistan

To discover the pollinator community of canola (Brassica napus L.) and the best pollinators for canola production, an experiment was performed at the research farm of Bahauddin Zakariya University, Multan, Pakistan. The insect pollinator community was composed of 35 species in 3 orders and 14 families. Most of the bees (Hymenoptera) and a butterfly species (Lepidoptera) foraged for nectar, whereas all the flies (Diptera) foraged either for pollen or both nectar and pollen. Eight major pollinators were tested for their pollination efficiency. The nectar-robbing behavior of many species made it difficult to judge the efficiency of an insect on the basis of visitation rate and stay time; therefore, the amounts of pollen deposited and pollen harvested per visit were also measured. The single visit efficiency in terms of the number of seeds per pod revealed that Apis dorsata, A. florea, and Halictus sp. were superior for canola pollination, having Spears’ values of 1.62, 1.55 and 1.73, respectively. With the increase in the number of seeds per pod, seed weight per pod also increased, confirming the importance of these three pollinator species in canola production.     

Specialty oilseed crops provide an abundant source of pollen for pollinators and beneficial insects

The continuing pollinator crisis is due, in part, to the lack of year‐round floral resources. In intensive farming regions, such as the Upper Midwest (UMW) of the USA, natural and pastoral vegetation largely has been replaced by annual crops such as maize (Zea mays L.), soyabean (Glycine max L.) and wheat (Triticum spp.). Neither the energy (nectar) nor protein (pollen) needs of pollinating and other beneficial insects are being met sufficiently by the new, high‐intensity, agricultural landscape. Several potentially useful oilseed crops can be grown in the UMW, and many of these oilseeds are highly attractive to beneficial insects. Prior research showed that some of these oilseeds produced abundant nectar, but their corresponding values for pollen production are unknown. Accordingly, the aim of our research was to document pollen (and protein) production per unit area of twelve oilseed crops grown in Minnesota and associate these values with levels of beneficial insect visitation during anthesis. Our results show that oilseed crops such as camelina (Camelina sativa L.), flax (Linum usitatissimum L.) and pennycress (Thlaspi arvense L.) produce relatively little pollen (≤40 kg/ha); borage (Borago officinalis L.), calendula (Calendula officinalis L.), canola (Brassica napus L.), crambe (Crambe abyssianica Hochst) and cuphea (Cuphea viscosissima Jacq. × Cuphea lanceolata W. T. Aiton) produce bountiful pollen resources (50–150 kg/ha); and oilseed echium (Echium plantagineum L.) generates massive amounts of pollen (>400 kg/ha), about 50% of which is protein. Our study is unique in presenting a season‐long perspective of pollen production in alternative oilseed crops, a resource valuable to pollen‐feeding insects such as managed and wild bees. Diversification of UMW landscapes that includes alternative oilseed crops such as oilseed echium and cuphea can potentially provide a ready source of pollen and protein to help combat pollinator decline.     

Norditerpene alkaloid concentrations in tissues and floral rewards of larkspurs and impacts on pollinators

Plant secondary compounds mediate interactions with insects and other animals. The norditerpene alkaloids are significant secondary compounds in Delphinium (larkspur) species which are divided into two classes: the 7, 8-methylenedioxylycoctonine (MDL-type) and N-(methylsuccinimido) anthranoyllycoctonine (MSAL-type), and are known to be toxic to herbivorous insects and livestock. Alkaloid concentrations were measured in a whole plant context in vegetative and floral tissues as well as rewards (pollen and nectar) in Delphinium barbeyi and Delphinium nuttallianum. Alkaloid concentrations differed between vegetative tissues, floral tissues and floral rewards. Alkaloid concentrations in floral parts were consistent with optimal defense theory, with tissues more closely tied to plant fitness, such as fruits, being more heavily defended than foliage. However, alkaloid concentrations were significantly lower in nectar compared to other tissues. The norditerpene alkaloids influenced the activity of bumble bees, the dominant pollinator of larkspur, but the effects were concentration dependent. Alkaloids in nectar are found at concentrations that have no effect on bee activity; however, if alkaloid concentrations in nectar were similar to those in foliage bee activity would be reduced significantly. These results suggest that nectar with low alkaloid concentrations may be beneficial to plant fitness by limiting adverse effects on pollinator activity.     

Acute exposure to a sublethal dose of imidacloprid and coumaphos enhances olfactory learning and memory in the honeybee Apis mellifera

The decline of honeybees and other pollinating insects is a current cause for concern. A major factor implicated in their decline is exposure to agricultural chemicals, in particular the neonicotinoid insecticides such as imidacloprid. Honeybees are also subjected to additional chemical exposure when beekeepers treat hives with acaricides to combat the mite Varroa destructor. Here, we assess the effects of acute sublethal doses of the neonicotinoid imidacloprid, and the organophosphate acaricide coumaphos, on honey bee learning and memory. Imidacloprid had little effect on performance in a six-trial olfactory conditioning assay, while coumaphos caused a modest impairment. We report a surprising lack of additive adverse effects when both compounds were administered simultaneously, which instead produced a modest improvement in learning and memory.     

Phyllotreta cruciferae and Phyllotreta striolata responses to insecticidal seed treatments with different modes of action

The strategy used most commonly in western North America to protect seedlings of canola (Brassica rapa L. and Brassica napus L.) from attack by adults of the flea beetles Phyllotreta cruciferae (Goeze) and Phyllotreta striolata (Fabricius) (Coleoptera: Chrysomelidae) involves planting seed coated with insecticide for systemic activity. Previous research determined that the two beetle species responded differently to the most commonly used neonicotinoid seed dressings. However, other insecticides that exploit different modes of action have commercial potential for managing infestations of these pests, but no information exists on their efficacies for these flea beetle species. Studies were conducted to compare effects of the neonicotinoid compounds, thiamethoxam and imidacloprid, to spinosyn and fipronil as systemic seed treatments for reducing feeding damage to canola seedlings and increasing mortality of P. cruciferae and P. striolata. Phyllotreta cruciferae experienced greater mortality and caused less feeding damage than P. striolata to seedlings treated with the neonicotinoid compounds. Mortality increased and feeding damage decreased significantly when beetles fed upon seedlings treated with fipronil, indicating its potential usefulness for control of these pests. However, spinosyn seed treatment was relatively ineffective against either beetle species. Higher rates of P. striolata mortality with fipronil than thiamethoxam suggest that fipronil may provide improved flea beetle control over hundreds of thousands of hectares in western North America where flea beetle populations are dominated by P. striolata, and control with thiamethoxam has been suboptimal.     

Adaptation for rodent pollination in Leucospermum arenarium (Proteaceae) despite rapid pollen loss during grooming

Background and Aims

Plants are adapted for rodent pollination in diverse and intricate ways. This study explores an extraordinary example of these adaptations in the pincushion Leucospermum arenarium (Proteaceae) from South Africa.

Methods

Live trapping and differential exclusion experiments were used to test the role of rodents versus birds and insects as pollinators. To explore the adaptive significance of geoflory, inflorescences were raised above ground level and seed production was compared. Captive rodents and flowers with artificial stigmas were used to test the effect of grooming on the rate of pollen loss. Microscopy, nectar composition analysis and manipulative experiments were used to investigate the bizarre nectar production and transport system.

Key Results

Differential exclusion of rodents, birds and insects demonstrated the importance of rodents in promoting seed production. Live trapping revealed that hairy-footed gerbils, Gerbillurus paeba, and striped field mice, Rhabdomys pumilio, both carried L. arenarium pollen on their forehead and rostrum, but much larger quantities ended up in faeces as a result of grooming. Terrarium experiments showed that grooming exponentially diminished the pollen loads that they carried. The nectar of L. arenarium was found to be unusually viscous and to be presented in a novel location on the petal tips, where rodents could access it without destroying the flowers. Nectar was produced inside the perianth, but was translocated to the petal tips via capillary ducts. In common with many other rodent-pollinated plants, the flowers are presented at ground level, but when raised to higher positions seed production was not reduced, indicating that selection through female function does not drive the evolution of geoflory.

Conclusions

Despite the apparent cost of pollen lost to grooming, L. arenarium has evolved remarkable adaptations for rodent pollination and provides the first case of this pollination system in the genus.     

Differential sensitivity of honey bees and bumble bees to a dietary insecticide (imidacloprid)

Currently, there is concern about declining bee populations and the sustainability of pollination services. One potential threat to bees is the unintended impact of systemic insecticides, which are ingested by bees in the nectar and pollen from flowers of treated crops. To establish whether imidacloprid, a systemic neonicotinoid and insect neurotoxin, harms individual bees when ingested at environmentally realistic levels, we exposed adult worker bumble bees, Bombus terrestris L. (Hymenoptera: Apidae), and honey bees, Apis mellifera L. (Hymenoptera: Apidae), to dietary imidacloprid in feeder syrup at dosages between 0.08 and 125 μg l^?1. Honey bees showed no response to dietary imidacloprid on any variable that we measured (feeding, locomotion and longevity). In contrast, bumble bees progressively developed over time a dose-dependent reduction in feeding rate with declines of 10–30% in the environmentally relevant range of up to 10 μg l^?1, but neither their locomotory activity nor longevity varied with diet. To explain their differential sensitivity, we speculate that honey bees are better pre-adapted than bumble bees to feed on nectars containing synthetic alkaloids, such as imidacloprid, by virtue of their ancestral adaptation to tropical nectars in which natural alkaloids are prevalent. We emphasise that our study does not suggest that honey bee colonies are invulnerable to dietary imidacloprid under field conditions, but our findings do raise new concern about the impact of agricultural neonicotinoids on wild bumble bee populations.     

Nutritional ecology of a parasitic wasp: food source affects gustatory response, metabolic utilization, and survivorship

The success of biological control is partly mediated by the longevity and reproductive success of beneficial insects. Availability of nectar and honeydew can improve the nutrition of parasitic insects, and thereby increase their longevity and realized fecundity. The egg parasitoid, Anaphes iole, showed strong gustatory perception of trehalulose, a carbohydrate found in homopteran honeydew. Chromatographic analysis demonstrated that enzymatic hydrolysis of sucrose, a common nectar sugar, proceeded at a faster rate than that of melezitose, a sugar common in aphid honeydew. A long-term bioassay showed that longevity was greater at 20 °C than at 27 °C, and at both temperatures survival was generally greatest for wasps provisioned with the three major nectar sugars, sucrose, glucose, and fructose. Patterns of food acceptance and utilization showed that A. iole accepted and utilized a broad range of sugars found in nature, including those found in nectar as well as honeydew. Glucose, fructose, and several oligosaccharides composed of these monosaccharide units appear to be more suitable for A. iole than other sugars tested. Evidence suggests that individual fitness benefits afforded by food sources are important for a time-limited parasitoid, and that continued investigations on the interface between nutrition and biological control are warranted for A. iole.     

Imidacloprid-induced impairment of mushroom bodies and behavior of the native stingless bee Melipona quadrifasciata anthidioides

Declines in pollinator colonies represent a worldwide concern. The widespread use of agricultural pesticides is recognized as a potential cause of these declines. Previous studies have examined the effects of neonicotinoid insecticides such as imidacloprid on pollinator colonies, but these investigations have mainly focused on adult honey bees. Native stingless bees (Hymenoptera: Apidae: Meliponinae) are key pollinators in neotropical areas and are threatened with extinction due to deforestation and pesticide use. Few studies have directly investigated the effects of pesticides on these pollinators. Furthermore, the existing impact studies did not address the issue of larval ingestion of contaminated pollen and nectar, which could potentially have dire consequences for the colony. Here, we assessed the effects of imidacloprid ingestion by stingless bee larvae on their survival, development, neuromorphology and adult walking behavior. Increasing doses of imidacloprid were added to the diet provided to individual worker larvae of the stingless bee Melipona quadrifasciata anthidioides throughout their development. Survival rates above 50% were only observed at insecticide doses lower than 0.0056 μg active ingredient (a.i.)/bee. No sublethal effect on body mass or developmental time was observed in the surviving insects, but the pesticide treatment negatively affected the development of mushroom bodies in the brain and impaired the walking behavior of newly emerged adult workers. Therefore, stingless bee larvae are particularly susceptible to imidacloprid, as it caused both high mortality and sublethal effects that impaired brain development and compromised mobility at the young adult stage. These findings demonstrate the lethal effects of imidacloprid on native stingless bees and provide evidence of novel serious sublethal effects that may compromise colony survival. The ecological and economic importance of neotropical stingless bees as pollinators, their susceptibility to insecticides and the vulnerability of their larvae to insecticide exposure emphasize the importance of studying these species.     

Specialized pollination in the African milkweed Xysmalobium orbiculare: a key role for floral scent in the attraction of spider-hunting wasps

Specialized pollination by prey-hunting wasps is poorly documented in rewarding plants. Furthermore, the mechanisms of achieving specialization are not clear since flowers typically produce exposed nectar and have no morphological adaptations (such as long spurs) to exclude non-pollinating visitors. We investigated the pollination of Xysmalobium orbiculare and explored the functional roles of floral scent and nectar in attracting pollinators and deterring nectar robbers. Floral visitor observations showed that this milkweed is visited almost exclusively by pompilid wasps in the genus Hemipepsis. These wasps were the only insects to carry pollinia, and a cage experiment confirmed their effectiveness in removing and inserting pollinia on flowers. Hand-pollinations showed that plants are genetically self-incompatible and thus reliant on pollinators for seed set. Palatability experiments with honeybees showed that nectar is distasteful to non-pollinating insects and is therefore likely to play a functional role in deterring nectar thieves. Choice experiments in the field showed that the wasp pollinators are attracted primarily by floral scent rather than visual cues. Analysis of spectral reflectance of flowers revealed that flowers are dull colored and are unlikely to stand out from the background vegetation. We conclude that X. orbiculare is specialized for pollination by spider-hunting wasps in the genus Hemipepsis and utilizes floral scent to selectively attract its pollinators and unpalatable nectar to deter non-pollinating visitors.     

Insect nicotinic acetylcholine receptor gene families: from genetic model organism to vector,pest and beneficial species

Nicotinic acetylcholine receptors (nAChRs) mediate fast synaptic transmission in the insect nervous system and are targets of a major group of insecticides, the neonicotinoids. Analyses of genome sequences have shown that nAChR gene families remain compact in diverse insect species, when compared to their mammalian counterparts. Thus, Drosophila melanogaster and Anopheles gambiae each possess 10 nAChR genes while Apis mellifera has 11. Although these are among the smallest nAChR gene families known, receptor diversity can be considerably increased by alternative splicing and mRNA A-to-I editing, thereby generating species-specific subunit isoforms. In addition, each insect possesses at least one highly divergent nAChR subunit. Species-specific subunit diversification may offer promising targets for future rational design of insecticides that act on particular pests while sparing beneficial insects. Electrophysiological studies on cultured Drosophila cholinergic neurons show partial agonist actions of the neonicotinoid imidacloprid and super-agonist actions of another neonicotinoid, clothianidin, on native nAChRs. Recombinant hybrid heteromeric nAChRs comprising Drosophila Dα2 and a vertebrate β2 subunit have been instructive in mimicking such actions of imidacloprid and clothianidin. Unitary conductance measurements on native nAChRs indicate that more frequent openings of the largest conductance state may offer an explanation for the superagonist actions of clothianidin.     

Nectar replenishment and pollen receipt interact in their effects on seed production of Penstemon roseus

Resource supply and pollen delivery are often thought to equally limit seed production in animal-pollinated plants. At equilibrium, plants should show no response to experimental pollen supplementation because resources limit seed set above the current level of pollen attraction, while experimental reduction in pollen deposition below the equilibrium level would reduce seed set. The predicted equilibrium may be disrupted, however, if plants expend additional energy to replenish removed nectar. We investigated the combined effects of nectar removal and pollen delivery on female reproductive success of Penstemon roseus (Plantaginaceae), a hummingbird-pollinated plant that replenishes removed nectar. We first documented that the frequency of experimental nectar removal was correlated with total nectar secretion; and increased frequency of nectar removal resulted in increased female reproductive costs to the plant. Trade-offs between investing resources in nectar and investing resources in seeds were then investigated in two contrasting natural populations by removing nectar from flowers at increasing frequencies while simultaneously hand-pollinating flowers with increasing amounts of pollen. Seed set was lowest at low levels of pollen deposition, highest at medium-sized pollen loads, and intermediate when pollen loads were highest. At both sites, the frequency of nectar removal and pollen deposition had an interactive effect on seed production, in that intermediate levels of nectar removal result in the absolute highest seed set, but only at intermediate pollen loads. At high pollen loads, seed set was higher following little to no nectar removal, and at low pollen loads, all rates of nectar removal affected fecundity equally. Seed mass responded to nectar removal and pollination differently than did seed set. High levels of nectar removal and pollen delivery both lowered seed mass, with little interaction between main effects. Our findings are among the first to demonstrate that nectar replenishment costs and pollination intensity jointly affect seed production. This conflict between nectar replenishment costs and pollen-limiting factors results in trade-offs between pollinator attraction and seed provisioning. Thus, resource allocation towards nectar production should more often be considered in future studies of pollen limitation.     

Bouncy versus idles: On the different role of pollinators in the generalist Gentiana lutea L.

Generalist flowers are visited by a broad variety of insects that function as pollinators, occasional visitors and as pollen and/or nectar robbers. Moreover, among legitimate pollinators the pollination efficiency can be different. Nectar greatly affects visitor behaviour and fidelity to a certain species, influencing plant reproductive effort. In this study we have investigated a generalist system (Gentiana lutea L.), examining the role of flower visitors and quantifying the contribution of each pollinating taxon in three natural populations. In order to verify the level of generalization, we introduce an index of Pollinator Performance (PoP), based on insect visitation rate and cross-pollen transport efficiency. Our results confirm the high degree of pollinator-generalization of the study species. Nevertheless, flower visitors show various degree of pollinating performance, mainly defined by their sedentary versus dynamic behaviour. Sedentary insects enhance geitonogamous pollen transfer, which results in reduced seed set and pollen limitation. In particular, an unusual sluggish behaviour was observed in bumblebees feeding on nectar. The hexose-rich abundant nectar offered by G. lutea flowers is remarkably rich in proline and β-alanine amino acids: this composition presumably influences feeding choice and insect dynamism, likely exerting a narcotic effect on pollinators. The consequences on plant fitness are discussed in an evolutionary perspective.